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| | In Vitro Urease Inhibition Assay |
| Description: | The assay mixture, containing 50 μl (2 mg/ml) of enzyme and 100 μl of different concentration of the tested agents, was added to 850 μl of 25 mM urea and preincubated for 0.5 h in water bath at 37 °C. The urease reaction was stopped after 30 min incubation by the following procedure. After pre-incubation, 500 μl of phenol reagent (1% w/v phenol and 0.005% w/v sodium nitroprusside) and 500 μl of alkali reagent (1% w/v NaOH and 0.075% active chloride NaOCl) were added to100 μl of incubation mixture and kept at 37 °C for 30 min. The absorbance (A) was measured at 625 nm by using following equation A = kbc, where c is the concentration of solution (mol/L), b the Length of the UV cell [Golbabaei et al., Daru., 21(1):2; Akhtar et al., Chem. Biol. Drug Des., 84:92-98]. All experiments were performed in triplicate in a final volume of 1 ml, and AHA was used as a standard urease inhibitor. |
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